Tocopherol-human serum albumin nanoparticles enhance lapatinib delivery and overcome doxorubicin resistance in breast cancer.

Introduction: HER2, a tyrosine kinase receptor, is amplified in HER2-positive breast cancer, driving cell signaling and growth. Aim: This study aimed to combat multidrug resistance in Dox-insensitive breast adenocarcinoma by creating a nanoformulation therapy with a tyrosine kinase inhibitor. Methodology: Human serum albumin (HSA) was conjugated with α-D-tocopherol succinate to form nanoaggregates loaded with lapatinib (Lapa). Results: The resulting Lapa@HSA(VE) NPs were 117.2 nm in size and demonstrated IC50 values of 10.25 µg/ml on MCF7 (S) and 8.02 µg/ml on MCF7 (R) cell lines. Conclusion: Lapa@HSA(VE) NPs showed no hepatotoxicity, unlike free Lapa, as seen in acute toxicity studies in rats.

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Bioactive extract of Morchella esculenta ameliorates cyclophosphamide-induced mitochondrial dysfunction and cardiotoxicity by modulating KEAP1/NRF2 and pro-inflammatory genes expression.

Prevention of anticancer drugs-induced cardiotoxicity remains an imperative area of oncology research as it continues to be a major challenge in cancer chemotherapy. This study was undertaken to investigate the protective effect of methanol extract of Morchella esculenta (ME) against cyclophosphamide (CP)-induced cardiotoxicity. Myocardial damage was assessed by biochemical and histopathological methods. Proinflammatory cytokines gene expression was determined by RT-PCR analysis. To assess the mitochondrial dysfunction, TCA cycle and electron transport chain complexes enzymes activities were determined. Chemical finger print of ME was accomplished by HPTLC. CP (200 mg/kg) treated animals showed elevation in cardiac injury markers which was attenuated by ME (p < 0.05). CP-induced decline of antioxidant status and expression of nuclear factor erythroid 2-related factor 2 were restored by ME. CP-induced expression of NF-ĸB, IL1-ß, IL-6, TNF-α, COX-2 and iNOS (p < 0.05) was attenuated by ME (500 mg/kg). Bioactive compounds namely, 5-eicosapentaenoicacid (C20H30O2), 8-hydroxyoctadecadienoic acid (C18H32O3), 4,4-dipo-zetacarotene (C30H44), CynarosideA (C21H32O10) present in the extract might be responsible for cardioprotection. The findings reveal the protective effect of ME against CP-induced cardiomyopathy.

Lactobacillus rhamnosus (LR) ameliorates pulmonary and extrapulmonary acute respiratory distress syndrome (ARDS) via targeting neutrophils.

Pulmonary and extrapulmonary acute respiratory distress syndrome (ARDS) is a life-threatening respiratory failure associated with high mortality. Despite progress in our understanding of the pathological mechanism causing the crippling illness, there are currently no targeted pharmaceutical treatments available for it. Recent discoveries have emphasized the existence of a potential nexus between gut and lung health fueling novel approaches including probiotics for the treatment of ARDS. We thus investigated the prophylactic-potential of Lactobacillus rhamnosus-(LR) in lipopolysaccharide (LPS)-induced pulmonary and cecal ligation puncture (CLP) induced extrapulmonary ARDS mice. Our in-vivo findings revealed that pretreatment with LR significantly ameliorated vascular-permeability (edema) of the lungs via modulating the neutrophils along with significantly reducing the expression of inflammatory-cytokines in the BALF, lungs and serum in both pulmonary and extrapulmonary mice-models. Interestingly, our ex-vivo immunofluorescence and flow cytometric data suggested that mechanistically LR via short chain fatty acids (butyrate being the most potent and efficient in ameliorating the pathophysiology of both pulmonary and extra-pulmonary ARDS) targets the phagocytic and neutrophils extracellular traps (NETs) releasing potential of neutrophils. Moreover, our in-vivo data further corroborated our ex-vivo findings and suggested that butyrate exhibits enhanced potential in ameliorating the pathophysiology of ARDS via reducing the infiltration of neutrophils into the lungs. Altogether, our study establishes the prophylactic role of LR and its associated metabolites in the prevention and management of both pulmonary and extrapulmonary ARDS via targeting neutrophils.

Predicting Obsessive-Compulsive Disorder Events in Children and Adolescents in the Wild using a Wearable Biosensor (Wrist Angel): Protocol for the Analysis Plan of a Nonrandomized Pilot Study.

BACKGROUND: Physiological signals such as heart rate and electrodermal activity can provide insight into an individual's mental state, which are invaluable information for mental health care. Using recordings of physiological signals from wearable devices in the wild can facilitate objective monitoring of symptom severity and evaluation of treatment progress. OBJECTIVE: We designed a study to evaluate the feasibility of predicting obsessive-compulsive disorder (OCD) events from physiological signals recorded using wrist-worn devices in the wild. Here, we present an analysis plan for the study to document our a priori hypotheses and increase the robustness of the findings of our planned study. METHODS: In total, 18 children and adolescents aged between 8 and 16 years were included in this study. Nine outpatients with an OCD diagnosis were recruited from a child and adolescent mental health center. Nine youths without a psychiatric diagnosis were recruited from the catchment area. Patients completed a clinical interview to assess OCD severity, types of OCD, and number of OCD symptoms in the clinic. Participants wore a biosensor on their wrist for up to 8 weeks in their everyday lives. Patients were asked to press an event tag button on the biosensor when they were stressed by OCD symptoms. Participants without a psychiatric diagnosis were asked to press this button whenever they felt really scared. Before and after the 8-week observation period, participants wore the biosensor under controlled conditions of rest and stress in the clinic. Features are extracted from 4 different physiological signals within sliding windows to predict the distress event logged by participants during data collection. We will test the prediction models within participants across time and multiple participants. Model selection and estimation using 2-layer cross-validation are outlined for both scenarios. RESULTS: Participants were included between December 2021 and December 2022. Participants included 10 female and 8 male participants with an even sex distribution between groups. Patients were aged between 10 and 16 years, and adolescents without a psychiatric diagnosis were between the ages of 8 and 16 years. Most patients had moderate to moderate to severe OCD, except for 1 patient with mild OCD. CONCLUSIONS: The strength of the planned study is the investigation of predictions of OCD events in the wild. Major challenges of the study are the inherent noise of in-the-wild data and the lack of contextual knowledge associated with the recorded signals. This preregistered analysis plan discusses in detail how we plan to address these challenges and may help reduce interpretation bias of the upcoming results. If the obtained results from this study are promising, we will be closer to automated detection of OCD events outside of clinical experiments. This is an important tool for the assessment and treatment of OCD in youth. TRIAL REGISTRATION: ClinicalTrials.gov NCT05064527; https://clinicaltrials.gov/study/NCT05064527. INTERNATIONAL REGISTERED REPORT IDENTIFIER (IRRID): DERR1-10.2196/48571.

Synthetic lethal mutants in Escherichia coli define pathways necessary for survival with RNase H deficiency.

Ribonucleotides frequently contaminate DNA and, if not removed, cause genomic instability. Consequently, all organisms are equipped with RNase H enzymes to remove RNA-DNA hybrids (RDHs). Escherichia coli lacking RNase HI (rnhA) and RNase HII (rnhB) enzymes, the ∆rnhA ∆rnhB double mutant, accumulates RDHs in its DNA. These RDHs can convert into RNA-containing DNA lesions (R-lesions) of unclear nature that compromise genomic stability. The ∆rnhAB double mutant has severe phenotypes, like growth inhibition, replication stress, sensitivity to ultraviolet radiation, SOS induction, increased chromosomal fragmentation, and defects in nucleoid organization. In this study, we found that RNase HI deficiency also alters wild-type levels of DNA supercoiling. Despite these severe chromosomal complications, ∆rnhAB double mutant survives, suggesting that dedicated pathways operate to avoid or repair R-lesions. To identify these pathways, we systematically searched for mutants synthetic lethal (colethal) with the rnhAB defect using an unbiased color screen and a candidate gene approach. We identified both novel and previously reported rnhAB-colethal and -coinhibited mutants, characterized them, and sorted them into avoidance or repair pathways. These mutants operate in various parts of nucleic acid metabolism, including replication fork progression, R-loop prevention and removal, nucleoid organization, tRNA modification, recombinational repair, and chromosome-dimer resolution, demonstrating the pleiotropic nature of RNase H deficiency. IMPORTANCE Ribonucleotides (rNs) are structurally very similar to deoxyribonucleotides. Consequently, rN contamination of DNA is common and pervasive across all domains of life. Failure to remove rNs from DNA has severe consequences, and all organisms are equipped with RNase H enzymes to remove RNA-DNA hybrids. RNase H deficiency leads to complications in bacteria, yeast, and mouse, and diseases like progressive external ophthalmoplegia (mitochondrial defects in RNASEH1) and Aicardi-Goutières syndrome (defects in RNASEH2) in humans. Escherichia coli ∆rnhAB mutant, deficient in RNases H, has severe chromosomal complications. Despite substantial problems, nearly half of the mutant population survives. We have identified novel and previously confirmed pathways in various parts of nucleic acid metabolism that ensure survival with RNase H deficiency.

Predicting obsessive-compulsive disorder episodes in adolescents using a wearable biosensor-A wrist angel feasibility study.

Introduction: Obsessive-compulsive disorders (OCD) are marked by distress, negative emotions, mental processes and behaviors that are reflected in physiological signals such as heart rate, electrodermal activity, and skin temperature. Continuous monitoring of physiological signals associated with OCD symptoms may make measures of OCD more objective and facilitate close monitoring of prodromal symptoms, treatment progress and risk of relapse. Thus, we explored the feasibility of capturing OCD events in the real world using an unobtrusive wrist worn biosensor and machine learning models. Methods: Nine adolescents (ages 10-17 years) with mild to moderate-severe OCD were recruited from child and adolescent mental health services. Participants were asked to wear the biosensor in the lab during conditions of rest and exposure to OCD symptom-triggering stimuli and for up to 8 weeks in their everyday lives and register OCD events. We explored the relationships among physiological data, registered OCD events, age, OCD symptom severity and symptom types. In the machine learning models, we considered detection of OCD events as a binary classification problem. A nested cross-validation strategy with either random 10-folds, leave-one-subject-out, or leave-week(s)-out in both layers was used. We compared the performance of four models: logistic regression, random forest (RF), feedforward neural networks, and mixed-effect random forest (MERF). To explore the ability of the models to detect OCD events in new patients, we assessed the performance of participant-based generalized models. To explore the ability of models to detect OCD events in future, unseen data from the same patients, we compared the performance of temporal generalized models trained on multiple patients with personalized models trained on single patients. Results: Eight of the nine participants collected biosensor signals totaling 2, 405 h and registered 1, 639 OCD events. Better performance was obtained when generalizing across time compared to across patients. Generalized temporal models trained on multiple patients were found to perform better than personalized models trained on single patients. RF and MERF models outperformed the other models in terms of accuracy in all cross-validation strategies, reaching 70% accuracy in random and participant cross-validation. Conclusion: Our pilot results suggest that it is possible to detect OCD episodes in the everyday lives of adolescents using physiological signals captured with a wearable biosensor. Large scale studies are needed to train and test models capable of detecting and predicting episodes. Clinical trial registration: ClinicalTrials.gov: NCT05064527, registered October 1, 2021.

Associations Between the Severity of Obsessive-Compulsive Disorder and Vocal Features in Children and Adolescents: Protocol for a Statistical and Machine Learning Analysis.

BACKGROUND: Artificial intelligence tools have the potential to objectively identify youth in need of mental health care. Speech signals have shown promise as a source for predicting various psychiatric conditions and transdiagnostic symptoms. OBJECTIVE: We designed a study testing the association between obsessive-compulsive disorder (OCD) diagnosis and symptom severity on vocal features in children and adolescents. Here, we present an analysis plan and statistical report for the study to document our a priori hypotheses and increase the robustness of the findings of our planned study. METHODS: Audio recordings of clinical interviews of 47 children and adolescents with OCD and 17 children and adolescents without a psychiatric diagnosis will be analyzed. Youths were between 8 and 17 years old. We will test the effect of OCD diagnosis on computationally derived scores of vocal activation using ANOVA. To test the effect of OCD severity classifications on the same computationally derived vocal scores, we will perform a logistic regression. Finally, we will attempt to create an improved indicator of OCD severity by refining the model with more relevant labels. Models will be adjusted for age and gender. Model validation strategies are outlined. RESULTS: Simulated results are presented. The actual results using real data will be presented in future publications. CONCLUSIONS: A major strength of this study is that we will include age and gender in our models to increase classification accuracy. A major challenge is the suboptimal quality of the audio recordings, which are representative of in-the-wild data and a large body of recordings collected during other clinical trials. This preregistered analysis plan and statistical report will increase the validity of the interpretations of the upcoming results. INTERNATIONAL REGISTERED REPORT IDENTIFIER (IRRID): DERR1-10.2196/39613.

Bioactive Extract of Morel Mushroom, Morchella esculenta (Ascomycota) Attenuates Doxorubicin-Induced Oxidative Stress Leading to Myocardial Injury.

Doxorubicin (DOX) is an extensively used anticancer drug for chemotherapy. Cardiotoxicity induced by DOX is an impediment in its clinical use. The aim of the current study is to evaluate the effect of a bioactive extract of an excellently edible morel mushroom, Morchella esculenta (ME) to attenuate DOX - induced cardiotoxicity. Protective effect of ME against DOX-induced cytotoxicity was evaluated in vitro by MTT assay using H9C2 cardiomyoblast cells. Intracellular free radical generation and mitochondrial membrane damage caused by DOX were detected by DCF-DA and rhodamine-123 dyes. Elevation of activities of creatine kinase-MB, lactate dehydrogenase and troponin I level consequent to the administration of DOX were determined using diagnostic kits. Depletion of endogenous antioxidant levels in myocardium was determined by spectrophotometric assays. Cardiac tissue damage caused by DOX was assessed by histopathological examination. ME reduced cytotoxicity caused by DOX at concentrations of 150 and 200 µg (p < 0.05 and p < 0.01, respectively). Cardiac injury marker levels elevated by DOX were significantly down regulated by ME (p < 0.01). Endogenous antioxidants such as SOD, GPx, and GSH depleted by DOX administration were restored to almost normal level by ME. This indicated the effect of ME to ameliorate oxidative stress caused by DOX administration leading to myocardial injury. Histopathological observation supported the finding. Being an excellently edible mushroom, current study indicates the potential therapeutic use of M. esculenta to prevent DOX-induced cardiotoxicity. The findings also suggest the clinical use of this medicinal mushroom to prevent chemo drug-induced cardiotoxicity.

Distinct Potentially Adaptive Accumulation of Truncation Mutations in Salmonella enterica serovar Typhi and Salmonella enterica serovar Paratyphi A.

Gene inactivation through the accumulation of truncation (or premature stop codon) mutations is a common mode of evolution in bacteria. It is frequently believed to result from reductive evolutionary processes allowing purging of superfluous traits. However, several works have demonstrated that, similar to the occurrences of inactivating nonsynonymous (i.e., amino acid replacement) mutations under positive selection pressures, truncation mutations can also be adaptive where specific traits deleterious in particular environmental conditions need to be inactivated for survival. Here, we performed a comparative analysis of genome-wide accumulation of truncation mutations in Salmonella enterica serovar Typhi and Salmonella enterica serovar Paratyphi A. Considering the known convergent evolutionary trajectories in these two serovars, we expected a strong overlap of truncated genes in S. Typhi and S. Paratyphi A, emerging through either reductive or adaptive dynamics. However, we detected a distinct set of core truncated genes encoding different overrepresented functional clusters in each serovar. In 54% and 28% truncated genes in S. Typhi and S. Paratyphi A, respectively, inactivating mutations were acquired by only different subsets of isolates, instead of all isolates analyzed for that serovar. Importantly, 62% truncated genes (P < 0.001) in S. Typhi and S. Paratyphi A were also targeted by convergent amino acid mutations in different serovars, suggesting those genes to be under selection pressures. Our findings indicate significant presence of potentially adaptive truncation mutations in conjunction with the ones emerging due to reductive evolution. Further experimental and large-scale bioinformatic studies are necessary to better explore the impact of such adaptive footprints of truncation mutations in the evolution of bacterial virulence. IMPORTANCE Detecting the adaptive mutations leading to gene inactivation or loss of function is crucial for understanding their contribution in the evolution of bacterial virulence and antibiotic resistance. Such inactivating mutations, apart from being of nonsynonymous (i.e., amino acid replacement) nature, can also be truncation mutations, abruptly trimming the length of encoded proteins. Importantly, the notion of reductive evolutionary dynamics is primarily accepted toward the accumulation of truncation mutations. However, our case study on S. Typhi and S. Paratyphi A, two human-restricted systemically invasive pathogens exerting similar clinical manifestations, indicated that a significant proportion of truncation mutations emerge from positive selection pressures. The candidate genes from our study will enable directed functional assays for deciphering the adaptive role of truncation mutations in S. Typhi and S. Paratyphi A pathogenesis. Also, our genome-level analytical approach will pave the way to understand the contribution of truncation mutations in the adaptive evolution of other bacterial pathogens.

Enhancement by postfiltering for speech and audio coding in ad hoc sensor networks.

Enhancement algorithms for wireless acoustic sensor networks (WASNs) are indispensable with the increasing availability and usage of connected devices with microphones. Conventional spatial filtering approaches for enhancement in WASNs approximate quantization noise with an additive Gaussian distribution, which limits performance due to the non-linear nature of quantization noise at lower bitrates. This work proposes a postfilter for enhancement based on Bayesian statistics to obtain a multidevice signal estimate, which explicitly models the quantization noise. The experiments using perceptual signal-to-noise ratio, perceptual evaluation of speech quality, and MUSHRA (multistimulus with hidden reference and anchors) scores demonstrate that the proposed postfilter can be used to enhance signal quality in ad hoc sensor networks.

Development of the Indian scale of the fear of COVID-19.

BACKGROUND: The first human case of COVID-19, the disease caused by the novel coronavirus causing COVID-19, subsequently named SARS-CoV-2 was reported by the officials in Wuhan City, China, in December 2019. In India, on January 31, 2020, the first case of COVID-19 was reported. MATERIALS AND METHODS: The Indian scale of fear related to COVID-19 (ISF-C19) Scale was developed in Hindi and was rated by two raters. Then, it was applied on twenty individuals (ten males and ten females) as a pilot study. The study had been conducted in Eastern India, in the state of Jharkhand. ISF-C19 was completed by 118 participants (females - 75; males - 43), aged 18 years or older from the community, and the subjective well-being was assessed. The psychometric properties of this instrument were investigated. Safety measures (i.e., mask, maintaining distance, gloves, and sanitizers) were taken throughout the data collection period. RESULTS: Findings suggested that this scale has adequate sampling, adequate interitem reliability coefficient, and higher overall scores on the ISF-C19 indicated more severe fear related to COVID-19. CONCLUSION: In light of the current scenario of the emerging cases in India, there is an urgent need to develop a scale related to COVID-19 as there is no published Indian standardized scale before this recent outbreak of pandemic. Considering the current scenario, this scale is useful to assess the fear related to COVID-19.

Chemical composition of ethanol extract of Macrotyloma uniflorum (Lam.) Verdc. using GC-MS spectroscopy.

BACKGROUND: Macrotyloma uniflorum Linn (Fabaceae) is a herbaceous plant with annual branches. It is used in kidney stones, inflamed joints, fever, musculoskeletal disorders, sinus wounds and localized abdominal tumors. It is reported as an antioxidant and nutraceutical (forage and food). GC-MS analysis of ethanol extract has led to identification of twenty-eight compounds from M. uniflorum by comparison of their retention indices and mass spectra fragmentation patterns with those stored on the GC-MS computer library. RESULTS: The main constituents identified were mome inositol, ethyl alpha-d-glucopyranoside, n- hexadecanoic acid, linoleic acid (9, 12-octadecadienoic acid), its esters and ethyl derivatives, Vitamin E, stigmasterol and 3-beta-stigmast-5-en-3-ol. CONCLUSIONS: The extracts are rich in linoleic acid and its esters, mome inositol and ethyl alpha-d-glucopyranoside; therefore, this plant can be medicinally beneficial as an antioxidant, in diabetes and its related disorders.

Lys-413 of S-phase mRNA cycling sequence binding protein from Leishmania donovani (LdCSBP) is modified through monoubiquitination that is responsible for inhibition of its riboendonuclease activity.

In addition to well-known process of proteasome-mediated degradation of polyubiquitinated proteins, monoubiquitination of proteins is also an important post-translational modification that regulates various non-degradative cellular processes like protein trafficking, cellular signalling, DNA replication and DNA repair. We have previously characterized a multi-domain cycling sequence binding protein LdCSBP from Leishmania donovani, which binds specifically to a conserved CAUAGAAG octamer containing RNAs via its uniquely arranged CCCH type Zn-fingers and degrades them using its Smr endonuclease domain, indicative of its potential role in the turnover of the S-phase mRNAs. Remarkably, its riboendonuclease activity is inhibited due to the incorporation of a monoubiquitin residue in the ZnF domain, though the target Lys residue remains unknown. Here, we report through systematic mutation of Lys residue to Ala that Lys-413 in LdCSBP is the site of monoubiquitination. However, the amino acid motif around the target Lys in LdCSBP is not consensus with any previously known monoubiquitination site, though partial homology is observed with a subset of recently identified mammalian ubiquitination target sites. Interestingly, Lys-413 of LdCSBP is conserved in the homologous annotated proteins from the related kinetoplastida parasites, suggesting similar monoubiquitination-mediated regulation of RNA endonuclease activity in the organisms.